Auswirkungen Konservierungsmittel-freier Fluorchinolon-Augenlösungen auf menschliche Hornhaut-Epithelzellen in vitro

2015 ◽  
Vol 1 (1) ◽  
pp. 26-27
Keyword(s):  
Scratch Wound ◽  
Scratch Wound Assay

Ziele: Die Beurteilung der biologischen Wirkungen Konservierungsmittel-freier Fluorchinolon-Augenlösungen auf Zellkulturen menschlichen Hornhaut-Epithels in vitro.Methoden: Wir untersuchten die Auswirkung von topischen Fluorchinolonen verschiedener Generationen, wie Ofloxacin 0,3%, Levofloxacin 0,5%, Tosufloxacin 0,3%, Moxifloxacin 0,5% und Gatifloxacin 0,3%, auf gezüchtete menschliche Hornhaut-Epithelzellen. Die Untersuchung erfolgte mittels MTT-basiertem kalorimetrischem Assay, Laktatdehydrogenase(LDH)-Leakage-Assay und Scratch-Wound-Assay. Die Morphologie der Hornhaut-Epithelzellen wurde mittels inverser Lichtmikroskopie und Transmissionselektronenmikroskopie untersucht.Ergebnisse: Bei allen topischen Fluorchinolonen ging die metabolische Aktivität der Hornhaut-Epithelzellen zeitabhängig zurück, und der LDH-Titer stieg mit zunehmender Dauer der Wirkstoffexposition. Insbesondere nach einer Exposition gegenüber Moxifloxacin 0,5% und Gatifloxacin 0,3% war ein signifikanter Anstieg der LDH-Titer im Vergleich zu den Kontrollen festzustellen. Die Migrationsraten der Hornhaut-Epithelzellen waren bei Ofloxacin 0,3% und Levofloxacin 0,5% höher als bei den anderen Fluorchinolonen. Nach einer Exposition gegenüber Moxifloxacin 0,5% und Gatifloxacin 0,3% waren schwere morphologische Schäden an den Zellen zu beobachten.Schlussfolgerung: Da Moxifloxacin 0,5% und Gatifloxacin 0,3% eine stärkere toxische Wirkung auf die Hornhaut-Epithelzellen ausübten als die anderen Fluorchinolone, sind diese Fluorchinolon-Augenlösungen der 4. Generation nur nach sorgfältiger Abwägung im Hinblick auf die mögliche Schädigung des Hornhaut-Epithels bei langer Behandlungsdauer oder zu hoher Dosierung anzuwenden.Übersetzung aus Ophthalmic Res 2014;51:216-223 (DOI: 10.1159/000357976)

scholarly journals Abnormal Cannabidiol Affects Production of Pro-Inflammatory Mediators and Astrocyte Wound Closure in Primary Astrocytic-Microglial Cocultures

Molecules ◽  
2020 ◽  
Vol 25 (3) ◽  
pp. 496 ◽  
Author(s):  
Julian Cardinal von Widdern ◽  
Tim Hohmann ◽  
Faramarz Dehghani
Keyword(s):  
Knockout Mice ◽  
Wound Closure ◽  
Scar Formation ◽  
Neuroprotective Effects ◽  
Proinflammatory Mediators ◽  
Nitrite Production ◽  
Scratch Wound ◽  
Scratch Wound Assay ◽  
The Impact

Abnormal cannabidiol (abn-CBD) exerts neuroprotective effects in vivo and in vitro. In the present study, we investigated the impact of abn-CBD on the glial production of proinflammatory mediators and scar formation within in vitro models. Primary astrocytic-microglial cocultures and astrocytic cultures from neonatal C57BL/6 mice and CB2 receptor knockout mice were stimulated with lipopolysaccharide (LPS), and the concentrations of tumor necrosis factor α (TNFα), interleukin-6 (IL-6) and nitrite were determined. Furthermore, we performed a live cell microscopy-based scratch-wound assay. After LPS stimulation, TNFα, IL-6 and nitrite production was more strongly increased in cocultures than in isolated astrocytes. Abn-CBD treatment attenuated the LPS-induced production of TNFα and nitrite in cocultures, while IL-6 production remained unaltered. In isolated astrocytes, only LPS-induced TNFα production was reduced by abn-CBD. Similar effects were observed after abn-CBD application in cocultures of CB2 knockout mice. Interestingly, LPS-induced TNFα and nitrite levels were far lower in CB2 knockout cultures compared to wildtypes, while IL-6 levels did not differ. In the scratch-wound assay, treatment with abn-CBD decelerated wound closure when microglial cells were present. Our data shows a differential role of abn-CBD for modulation of glial inflammation and astrocytic scar formation. These findings provide new explanations for mechanisms behind the neuroprotective potential of abn-CBD.

Effect of different chitosan derivatives on in vitro scratch wound assay: A comparative study

2015 ◽  
Vol 76 ◽  
pp. 236-241 ◽  
Author(s):  
Francesca Felice ◽  
Ylenia Zambito ◽  
Ester Belardinelli ◽  
Angela Fabiano ◽  
Tatiana Santoni ◽  
...  
Keyword(s):  
Comparative Study ◽  
Chitosan Derivatives ◽  
Scratch Wound ◽  
Scratch Wound Assay

Synthesis and evaluation of an alginate-methacrylate xerogel for insulin delivery towards wound healing applications

2021 ◽  
Vol 12 (3) ◽  
pp. 215-234
Author(s):  
GP Rajalekshmy ◽  
MR Rekha
Keyword(s):  
Wound Healing ◽  
Transmission Rate ◽  
Wound Care ◽  
Physical Stability ◽  
Analytical Techniques ◽  
Insulin Delivery ◽  
Water Vapour Transmission Rate ◽  
Scratch Wound ◽  
Scratch Wound Assay

Background: Alginate is one of the most widely used biopolymer for wound healing. But poor mechanical strength and degradability limits its application especially as a drug-delivery matrix. The aim of this study was to develop stable alginate based scaffold for insulin delivery toward wound care. Materials & methods: The xerogel alginate-g-poly (methacrylic acid; AGM2S) was characterized by various analytical techniques. Results: AGM2S xerogel showed improved physical stability, low degradation, good swelling and water vapour transmission rate (WVTR). About 70% of insulin was released from loaded xerogel over a period of 48 h and favorably modulated the healing response in in vitro scratch wound assay. Conclusion: Grafting improved the strength and stability of alginate xerogel and the results suggest the application of insulin loaded AGM2S xerogels as a potential wound healing material.

scholarly journals Avaliação dos efeitos do inibidor multiquinase sorafenibe sobre as células musculares lisas arteriais de Rattus norvegicus: Estudo in vitro / Evaluation of the effects of the multikinase inhibitor sorafenib on the arterial smooth muscle cells of Rattus norvegicus: An in vitro study

2021 ◽  
Vol 7 (12) ◽  
pp. 116307-116318
Author(s):  
Rafael de Nogueira Riberio ◽  
Gabriela Mendonça Dos Reis ◽  
Laura Bainy Rodrigues De Freitas ◽  
Gabriela Jouglard Vasquez Amado ◽  
Jessica Machado Miri ◽  
...  
Keyword(s):  
Smooth Muscle ◽  
Smooth Muscle Cells ◽  
Rattus Norvegicus ◽  
In Vitro Study ◽  
Multikinase Inhibitor ◽  
Scratch Wound ◽  
Scratch Wound Assay ◽  
Ciclo Celular ◽  
Arterial Smooth Muscle Cells

A reestenose arterial é um processo inflamatório que pode ocorrer após colocação de stent por cateterismo. Os stents farmacológicos surgiram para reduzir esse problema e o inibidor multiquinase sorafenibe demonstrou ser um composto com ação efetiva. Este estudo in vitro avaliou os efeitos do sorafenibe sobre a citotoxicidade, migração celular e distribuição das células nas fases do ciclo celular. A linhagem celular de músculo liso de rato A7r5 foi tratada com sorafenibe em concentrações que variaram de 0 a 5 μM. Os efeitos citotóxicos foram avaliados por dois ensaios colorimétricos, MTT e SRB após 24 horas de tratamento. A distribuição das células nas fases do ciclo celular foi avaliada por citometria de fluxo e a capacidade de cicatrização/migração celular pelo ensaio scratch wound assay. Comparado com o controle positivo paclitaxel, o sorafenibe demonstrou um efeito 1,6 vezes maior na redução da proliferação celular. Na avaliação do ciclo celular, o sorafenibe mostrou um bloqueio na fase G0/G1. Além disso, o sorafenibe aumentou o número de A7r5 células na fase sub-G1, sugerindo morte celular. No entanto, no estudo de cicatrização/migração celular, não foi observado efeito quando comparado ao controle negativo. Assim, esses resultados in vitro sugerem que o sorafenibe é eficaz para uso em stents farmacológicos, sugerindo uma continuidade na investigação desse fármaco.

scholarly journals Paracrine Proangiogenic Function of Human Bone Marrow-Derived Mesenchymal Stem Cells Is Not Affected by Chronic Kidney Disease

2019 ◽  
Vol 2019 ◽  
pp. 1-12 ◽  
Author(s):  
Femke C. C. van Rhijn-Brouwer ◽  
Bas W. M. van Balkom ◽  
Diana A. Papazova ◽  
Diënty H. M. Hazenbrink ◽  
Anke J. Meijer ◽  
...  
Keyword(s):  
Chronic Kidney Disease ◽  
Bone Marrow ◽  
Kidney Disease ◽  
Kidney Transplant ◽  
Galactosidase Activity ◽  
Kidney Transplant Recipients ◽  
Paracrine Factors ◽  
Differentiation Capacity ◽  
Scratch Wound

Background. Cell-based therapies are being developed to meet the need for curative therapy in chronic kidney disease (CKD). Bone marrow- (BM-) derived mesenchymal stromal cells (MSCs) enhance tissue repair and induce neoangiogenesis through paracrine action of secreted proteins and extracellular vesicles (EVs). Administration of allogeneic BM MSCs is less desirable in a patient population likely to require a kidney transplant, but potency of autologous MSCs should be confirmed, given previous indications that CKD-induced dysfunction is present. While the immunomodulatory capacity of CKD BM MSCs has been established, it is unknown whether CKD affects wound healing and angiogenic potential of MSC-derived CM and EVs. Methods. MSCs were cultured from BM obtained from kidney transplant recipients (N=15) or kidney donors (N=17). Passage 3 BM MSCs and BM MSC-conditioned medium (CM) were used for experiments. EVs were isolated from CM by differential ultracentrifugation. BM MSC differentiation capacity, proliferation, and senescence-associated β-galactosidase activity was assessed. In vitro promigratory and proangiogenic capacity of BM MSC-derived CM and EVs was assessed using an in vitro scratch wound assay and Matrigel angiogenesis assay. Results. Healthy and CKD BM MSCs exhibited similar differentiation capacity, proliferation, and senescence-associated β-galactosidase activity. Scratch wound migration was not significantly different between healthy and CKD MSCs (P=0.18). Healthy and CKD BM MSC-derived CM induced similar tubule formation (P=0.21). There was also no difference in paracrine regenerative function of EVs (scratch wound: P=0.6; tubulogenesis: P=0.46). Conclusions. Our results indicate that MSCs have an intrinsic capacity to produce proangiogenic paracrine factors, including EVs, which is not affected by donor health status regarding CKD. This suggests that autologous MSC-based therapy is a viable option in CKD.

scholarly journals The Activity of Combination of Hydrolyzed Virgin Coconut Oil and Chitosan Toward Wound Healing Parameters on NIH 3T3 Cells Using in Vitro Methods

1970 ◽  
Vol 7 (3) ◽  
pp. 14-19 ◽  
Author(s):  
Hekdin Marsius Sipayung ◽  
Jansen Silalahi ◽  
Yuandani Y
Keyword(s):  
Cell Proliferation ◽  
Wound Healing ◽  
Protein Expression ◽  
Scratch Wound ◽  
Cell Proliferation Activity ◽  
Proliferation Activity ◽  
Cox 2 ◽  
Nih 3T3 ◽  
Vegf Protein

Objectives: The objective of this study was to investigate the activity of combination of hydrolyzed VCO (HVCO) and chitosan on NIH 3T3 cell proliferation activity, NIH 3T3 cell migration, COX-2 and VEGF protein expression. Design: In vitro cytotoxic assay was determined by MTT (MicrocultureTetrazoliumTehnique) assay, cell proliferation activity was measured by calculating cell viability incubated 24 hours, 48 hours and 72 hours, wound closure percentage was tested by scratch wound healing method, expression of COX-2 protein and VEGF protein were measured by immunocytochemical method. Interventions: The variable that was intervened in this study was the concentration of HVCO and chitosan. Main Outcome Measures: The main measurements carried out in this study were the absorbance value of HVCO and chitosan which was converted into viability cell, proliferation activity, percentage of wound closure, and percentage of COX-2 and VEGF protein expression. Results: Cytotoxic activity of HVCO and chitosan resulted the best concentration at 31.25 μg/ml, scratch wound healing assay from a combination HVCO and chitosan resulted the best migration of fibroblast cells at a ratio of 1:1 with HVCO 62.5 μg/ml and chitosan 62.5 μg/ml, combination of HVCO 62.5 μg/ml and chitosan 62.5 μg/ml (1:1) increased expression of COX-2 and VEGF. Conclusion: Combination of HVCO and chitosan could increase NIH 3T3 cell migration, COX-2 and VEGF protein expression. Combination of HVCO and chitosan had better wound healing activity in vitro than single use. Keywords: Rhizomucor miehei, viability, proliferation, migration, expression

scholarly journals An image J plugin for the high throughput image analysis of in vitro scratch wound healing assays

2020 ◽  
Author(s):  
Alejandra Suarez-Arnedo ◽  
Felipe Torres Figueroa ◽  
Camila Clavijo ◽  
Pablo Arbeláez ◽  
Juan C. Cruz ◽  
...  
Keyword(s):  
Wound Healing ◽  
Image Analysis ◽  
Low Cost ◽  
Cellular Migration ◽  
Wound Healing Assay ◽  
Wound Area ◽  
Scratch Wound ◽  
Scratch Wound Healing Assay ◽  
Imagej Plugin

AbstractIn vitro scratch wound healing assay, a simple and low-cost technique that works along with other image analysis tools, is one of the most widely used 2D methods to determine the cellular migration and proliferation in processes such as regeneration and disease. There are open-source programs such as imageJ to analyze images of in vitro scratch wound healing assays, but these tools require manual tuning of various parameters, which is time-consuming and limits image throughput. For that reason, we developed an optimized plugin for imageJ to automatically recognize the wound healing size, correct the average wound width by considering its inclination, and quantify other important parameters such as: area, wound area fraction, average wound width, and width deviation of the wound images obtained from a scratch/ wound healing assay. Our plugin is easy to install and can be used with different operating systems. It can be adapted to analyze both individual images and stacks. Additionally, it allows the analysis of images obtained from bright field, phase contrast, and fluorescence microscopes. In conclusion, this new imageJ plugin is a robust tool to automatically standardize and facilitate quantification of different in vitro wound parameters with high accuracy compared with other tools and manual identification.

Dissecting the invasive potential of esophageal cancer by siRNA library screening.

2015 ◽  
Vol 33 (3_suppl) ◽  
pp. 86-86
Author(s):  
Shane P Duggan ◽  
Sinead Phipps ◽  
Fiona Behan ◽  
Catherine Garry ◽  
Dermot P Kelleher
Keyword(s):  
Cell Proliferation ◽  
Esophageal Cancer ◽  
Cell Viability ◽  
Metastatic Potential ◽  
Software Systems ◽  
Invasive Potential ◽  
Significant Difference ◽  
Scratch Wound ◽  
Scratch Wound Assay ◽  
Reverse Transfection

86 Background: The low survival for esophageal cancer is in part attributed to its high invasive potential and distant metastasis. In cancer, abnormal cell migration is an essential component of metastasis, and it is reasonable to consider that the conversion between different forms of morphology permits tumor cells to metastasise. Discovering regulators of esophageal cancer morphogenesis may aid in the development of novel targeted therapies that limit metastatic potential. Methods: GOhTRT cells were seeded and treated with siRNA (Human Druggable Genome, Dharmacon) by reverse transfection. Cells were fixed, immunostained for DNA, tubulin and actin and imaged with automated microscope. Cell images were processed using the InCell Analyzer software and the R statistical software systems CellHTS2 and RNAither. The effect of RNAi knockdown on cell viability and cell motility were assessed using MTT cell proliferation assay and scratch wound assay. Results: 127 gene candidates greatly exhibited effects on F-actin and α-tubulin area staining. This list was refined to six high quality candidates (RRM2, ITGB8, GPS1, SPRY1, NOL1 and MYO9B). Silencing of GPS1, MYO9B and SPRY1 increased the rate of migration in a scratch wound assay, with 86.98% ± 3.097%, 75.78% ±5.454% and 72.97% ± 5.463% (p =0.0022) respectively. There was no significant difference in cell viability absorbance values for siGPS1 (0.9037 ± 0.06575; p =0.1905) and siSPRY1 (0.9088 ± 0.09849; p =0.2985), which suggests that the increased rate of wound closure previously seen is by virtue of migratory signalling as oppose to an increase in cell proliferation. Cell viability was decreased considerably in siRRM2 cells (0.2492 ± 0.02798; p <0.0001) and siMYO9B (0.4048 ± 0.04663; p<0.0001) in comparison to siNT cells (1.046 ± 0.07712). Conclusions: In summary, this screen successfully identified high confidence hits associated with cytoskeletal remodelling, some of which are already associated with metastasis in literature and database searches. Further mechanistic studies and gene pathway analysis of candidate genes will provide novel therapeutic targets which can be utilised to block the spread of cancer in patients.

scholarly journals Combination of Two Kinds of Medicated Microparticles Based on Hyaluronic Acid or Chitosan for a Wound Healing Spray Patch

Pharmaceutics ◽  
2021 ◽  
Vol 13 (12) ◽  
pp. 2195
Author(s):  
Angela Fabiano ◽  
Chiara Migone ◽  
Luca Cerri ◽  
Anna Maria Piras ◽  
Andrea Mezzetta ◽  
...  
Keyword(s):  
Wound Healing ◽  
Hyaluronic Acid ◽  
Second Life ◽  
Olive Leaves ◽  
Waste Products ◽  
Accelerate Wound Healing ◽  
Scratch Wound ◽  
Treatment Of Wounds ◽  
Scratch Wound Healing Assay

Olive leaves extract (OLE) has been extensively studied as antioxidant and antibiotic and these characteristics make it particularly interesting for use on wounds. For this reason, the aim of this study was to introduce OLE in microparticles (MP) of hyaluronic acid (MPHA-OLE) or chitosan (MPCs-OLE) to obtain a spray patch for the treatment of wounds in anatomical areas that are difficult to protect with traditional patches. The MP were characterized for particle size and ability to protect OLE from degradation, to absorb water from wound exudate, to control OLE release from MP. The MPHA and MPCs medicated or not and mixtures of the two types in different proportions were studied in vitro on fibroblasts by the scratch wound healing assay. The MP size was always less than 5 µm, and therefore, suitable for a spray patch. The MPCs-OLE could slow down the release of OLE therefore only about 60% of the polyphenols contained in it were released after 4 h. Both MPHA and MPCs could accelerate wound healing. A 50% MPHA-OLE-50% MPCs-OLE blend was the most suitable for accelerating wound healing. The MPHA-OLE-MPCs-OLE blends studied in this work were shown to have the characteristics suitable for a spray patch, thus giving a second life to the waste products of olive growers.

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